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\title{\emph{In vivo} cleavage of solubility tags as a tool to enhance the
levels of soluble recombinant proteins in \emph{Escherichia coli}}



\author[1]{Filipe Silva}%
\author[1]{Sara Santos}%
\author[1]{Roberto Meyer}%
\author[1]{Eduardo Silva}%
\author[1]{Carina Pinheiro}%
\author[1]{Neuza Alcantara-Neves}%
\author[1]{Luis Pacheco}%
\affil[1]{Federal University of Bahia}%


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  \date{March 13, 2025}


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\begin{abstract}
Recombinant proteins are generally fused with solubility enhancer tags
to improve target protein folding and solubility. However, the fusion
protein strategy usually requires the use of expensive proteases to
perform in vitro proteolysis and additional chromatography steps to
obtain tag-free recombinant proteins. Expression systems based on
intracellular processing of solubility tags in Escherichia coli, through
co-expression of a site-specific protease, are useful for simplifying
the recombinant protein purification process, for screening molecules
that fail to remain soluble after tag removal, and to promote higher
yields of soluble target protein. Herein, we review controlled
intracellular processing (CIP) systems, tailored to produce soluble
untagged proteins in E. coli. We discuss the different genetic systems
available for intracellular protein processing regarding system design
features, significant advantages and limitations of the various
strategies.%
\end{abstract}%



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